过表达Notch1的胞内段基因对人牙周膜干细胞增殖能力影响

Effect of Over Expression of NICD Gene on Proliferation of Human Periodontal Ligament Stem Cells

目的:探讨过表达Notch1的胞内段(Notch1intracellular domain,NICD)基因对人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)增殖能力的影响。方法:构建含NICD基因过表达的逆转录病毒颗粒,并将其转染至牙周膜干细胞,构建过表达NICD的细胞株PDLSCs/NICD,转染48h后观察细胞一般状态。PDLSCs/NICD为实验组,以PDLSCs/wt为正常细胞组和PDLSCs/vector空病毒转染组作为对照,通过实时定量聚合酶链锁反应(quantitative Real-time-Polymerase chain reaction,qRT-PCR)和Western blot检测NICD mRNA及其蛋白的表达;细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞的增殖情况;Transwell实验检测细胞的迁移能力。结果:Western blot和qRT-PCR结果显示,与PDLSCs/wt组和PDLSCs/vector组比较,PDLSCs/NICD组的NICD蛋白表达水平及其mRNA表达水平均明显升高(P<0.05);CCK-8结果提示,与PDLSCs/vector组比较,PDLSCs/NICD组细胞增殖速度加快,Transwell实验结果显示,PDLSCs/NICD组的细胞迁移(40.20±1.74)明显高于PDLSCs/vector组(21.20±1.18)。结论:NICD基因过表达的人PDLSCs在一定程度上增殖及迁移能力加强。

Objective: To investigate the effect of Notch1 intracellular domain (NICD) gene on the proliferation of human periodontal ligament stem cells (PDLSCs).Methods: The retroviral particles with over expression of Notch1 (NICD) gene were constructed and transfected into the PDLSCs.The general state of the cells was observed after 48h transfection.PDLSCs/NICD was set as the experimental group,PDLSCs/wt was set as normal group,and PDLSCs/empty vector was set as control group.The expressions of NICD mRNA and protein were detected by Real-time PCR and Western Blot,the proliferation of cells was detected by CCK8 method,and the migration ability of the cells was detected.Results: The expression levels of protein and NICD mRNA in the PDLSCs/NICD group were obviously enhanced in contrast to PDLSCs/wt group and the PDLSCs/empty vector group.Cell migration of PDLSCs/NICD group (40.20±1.738) was significantly higher than that of PDLSCs/vector group (21.20± 1.182 ).Conclusion: Over expression of NICD enhanced the proliferation and migration ability of human PDLSCs to a certain extent.